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991.
Yu Ban Ying-ying Wu Tao Yu Ning Geng Yong-yue Wang Xiao-guang Liu Ping Gong 《Tissue & cell》2011,43(5):311-317
The process of mechanotransduction of bone, the conversion of a mechanical stimulus into a biochemical response, is known to occur in osteoblasts in response to fluid shear stress. In order to understand the reaction of osteoblasts to various times of flow perfusion, osteoblasts were seeded on three-dimensional scaffolds, and cultured in the following conditions: continuous flow perfusion, intermittent flow perfusion, and static condition. We collected samples on day 4, 8 and 12 for analysis. Osteoblast proliferation was demonstrated by cell proliferation and scanning electron microscopy assay. Additionally, the expression of known markers of differentiation, including alkaline phosphatase and osteocalcin, were tested by qRT-PCR and alkaline phosphatase activity assay, and the deposition of calcium was used as an indicator of mineralization demonstrated by calcium content assay. The results supported that low fluid shear stress plays an important role in the activation of osteoblasts: enhance cell proliferation, increase calcium deposition, and promote the expression of osteoblastic markers. Furthermore, the continuous flow perfusion is a more favorable environment for the initiation of osteoblast activity compared with intermittent flow perfusion. Therefore, the force and time of fluid shear stress are important parameters for osteoblast activation. 相似文献
992.
A Gram positive, rod-shaped potential strain was selected from the pool of bacterial isolates obtained from the Western Ghats
forest (India) on the basis of zone of P-solubilization activity. Identification based on 16S rRNA gene sequence revealed
that the strain is of Bacillus species, sharing highest sequence similarity to Bacillus tequilensis NRRL B-41771T (99.5%). Strain NII-0943 was able to produce good amount of indole acetic acid (IAA) and was positive for siderophore production.
In addition to IAA and siderophore attributes, strain NII-0943 also possessed the characteristics like Ca3(PO4)2 solubilization and growth in nitrogen-free medium. Seed inoculation with the strain NII-0943 resulted in significantly higher
root initiation in black pepper cuttings grown under pots. The contents of nitrogen and phosphorus in both soil and plant
were also enhanced significantly in treatments inoculated with these bacterial inocula. Hence, based on this evidence it is
proposed that strain NII-0943 could be deployed as a plant growth-promoting inoculant to attain the desired results of bacterization. 相似文献
993.
Xiao F Gao W Wang X Chen T 《Apoptosis : an international journal on programmed cell death》2012,17(6):600-611
Although caspases have been demonstrated to be involved in artemisinin (ARTE)-induced apoptosis, their exact functions are
not well understood. The aim of this report is to explore the roles of caspase-8, -9 and -3 during ARTE-induced apoptosis
in human lung adenocarcinoma (ASTC-a-1) cells. ARTE treatment induces a rapid generation of reactive oxygen species (ROS),
and ROS-dependent apoptosis as well as the activation of caspase-8, -9 and -3 via time- and dose-dependent fashion. Of upmost
importance, inhibition of caspase-8 or -9, but not caspase-3, almost completely blocks the ARTE-induced not only activation
of the caspase-8, -9 and -3 but also apoptosis. In addition, the apoptotic process triggered by ARTE does not involve the
Bid cleavage, tBid translocation, significant loss of mitochondrial membrane potential and cytochrome c release from mitochondria.
Moreover, silencing Bax/Bak does not prevent the ATRE-induced cell death as well as the activation of caspase-8, -9 and -3.
Collectively, our data firstly demonstrate that ARTE triggers a ROS-mediated positive feedback amplification activation loop
between caspase-8 and -9 independent of mitochondria, which dominantly mediated the ARTE-induced apoptosis via a caspase-3-independent
apoptotic pathway in ASTC-a-1 cells. Our findings imply a potential to develop new derivatives from artemisinin to effectively
initiate the amplification activation loop of caspases. 相似文献
994.
Rhamnolipids, produced by Pseudomonas aeruginosa, represent an important group of biosurfactants having various industrial, environmental, and medical applications. Current
methods for rhamnolipid quantification involve the use of strong hazardous acids/chemicals, indirect measurement of the concentration
of sugar moiety, or require the availability of expensive equipment (HPLC-MS). A safer, easier method that measures the whole
rhamnolipid molecules would significantly enhance strain selection, metabolic engineering, and process development for economical
rhamnolipid production. A semi-quantitative method was reported earlier to differentiate between the rhamnolipid-producing
and non-producing strains using agar plates containing methylene blue and cetyl trimethylammonium bromide (CTAB). In this
study, a rapid and simple method for rhamnolipid analysis was developed by systematically investigating the complexation of
rhamnolipids and methylene blue, with and without the presence of CTAB. The method relies on measuring the absorbance (at
638 nm) of the rhamnolipid−methylene blue complex that partitions into the chloroform phase. With P. aeruginosa fermentation samples, the applicability of this method was verified by comparison of the analysis results with those obtained
from the commonly used anthrone reaction technique. 相似文献
995.
There is an apparent allometric relationship between peak frequency of echolocation and body size in rhinophilids. However,
some rhinolophids deviate from this rule. To date this variation has been explained as a result of partitioning of communication
channels. An alternative hypothesis that food resource partitioning results in this divergence in expected frequencies was
tested by comparing prey selection between Rhinolophus macrotis Blyth, 1844 and Rhinolophus lepidus Blyth, 1844 in Yunnan Province, China. These two sympatric species are morphologically similar but acoustically divergent:
R. macrotis has an echolocation frequency significantly lower than that predicted by the allometric relationship, whereas that of R. lepidus agreed with expectations. Prey selection experiments, conducted in a flight tent, indicated that the dominant prey taxa of
R. macrotis were Lasiocampidae, Arctiidae and Noctuidae, whilst that of R. lepidus were Arctiidae, Noctuidae and Ichneumonidae. R. macrotis ate more earless moths and fewer eared moths than R. lepidus, and R. macrotis fed on larger prey in general and captured a wider size range than that captured by R. lepidus. These results confirmed the existence of finely tuned trophic niche differentiation and suggested that food resource partitioning
is one of the factors leading to lower peak frequency of calls in R. macrotis. 相似文献
996.
Xiaohuan Cheng Junfa Ding Fang Zheng Xin Zhou Chenling Xiong 《Molecular biology reports》2009,36(8):2053-2057
Familial hypercholesterolemia (FH) (OMIM 143890) is an autosomal dominantly inherited disease mainly caused by mutations of
the gene encoding the low density lipoprotein receptor (LDLR) and Apolipoprotein (Apo) B. First the common mutation R3500Q
in ApoB gene was determined using PCR/RFLP method. Then the LDLR gene was screened for mutations using Touch-down PCR, SSCP
and sequencing techniques. Furthermore, the secondary structure of the LDLR protein was predicted with ANTHEPROT5.0. The R3500Q
mutation was absent in these two families. A heterozygous p.W483X mutation of LDLR gene was identified in family A which caused
a premature stop codon, while a homozygous mutation p.A627T was found in family B. The predicted secondary structures of the
mutant LDLR were altered. We identified two known mutations (p.W483X, p.A627T) of the LDLR gene in two Chinese FH families
respectively. 相似文献
997.
可溶性肿瘤坏死因子受体II-脂联素球部融合蛋白的真核表达及生物学活性检测 总被引:1,自引:1,他引:1
本研究设计和构建了一种人肿瘤坏死因子受体II胞外区与人脂联素球部的融合基因sTNFRII-gAD,且相应的融合蛋白在哺乳动物细胞BHK-21S的无血清培养体系中实现了表达,并对该融合蛋白进行了初步鉴定。首先,用RT-PCR方法从人的外周血淋巴细胞总RNA中扩增人肿瘤坏死因子II型受体胞外区基因片段,与脂联素球部基因片段融合,克隆至pAAV2neo表达载体中,构建成pAAV2neo-sTNFRII-gAD。随后,用pAAV2neo-sTNFRII-gAD转染BHK-21S细胞获得G418抗性细胞BHK-21S/pAAV2neo-sTNFRII-gAD;然后,将原来含有血清的培养液换成无血清的化学成分限定的培养液,细胞从贴壁培养方式转换成悬浮培养方式;最后,收集BHK-21S/pAAV2neo-sTNFRII-gAD无血清悬浮培养24h后的培养上清,进行sTNFRII-gAD融合蛋白的鉴定分析。酶切鉴定和测序结果显示,所构建的pAAV2neo-sTNFRII-gAD质粒结构正确,sTNFRII-gAD序列与预期一致;分别用抗人肿瘤坏死因子受体II和抗人脂联素球部的单克隆抗体检测pAAV2neo-sTNFRII-gAD瞬时转染的BHK-21S细胞,免疫荧光呈现阳性;免疫印迹分析在pAAV2neo-sTNFRII-gAD稳定转染的BHK-21S细胞上清中检测到sTNFRII-gAD融合蛋白的表达,并以单体、三聚体和三聚体以上的多聚体形式存在。活性测定结果表明,sTNFRII-gAD融合蛋白具有显著抑制TNFα杀伤L929细胞的活性。因此,本研究为下一步大量制备sTNFRII-gAD融合蛋白用于体内外功能研究提供了良好基础。 相似文献
998.
999.
1000.
Systematic Evaluation of Protein Sequence Filtering Algorithms for Proteoform Identification Using Top‐Down Mass Spectrometry 下载免费PDF全文
Complex proteoforms contain various primary structural alterations resulting from variations in genes, RNA, and proteins. Top‐down mass spectrometry is commonly used for analyzing complex proteoforms because it provides whole sequence information of the proteoforms. Proteoform identification by top‐down mass spectral database search is a challenging computational problem because the types and/or locations of some alterations in target proteoforms are in general unknown. Although spectral alignment and mass graph alignment algorithms have been proposed for identifying proteoforms with unknown alterations, they are extremely slow to align millions of spectra against tens of thousands of protein sequences in high throughput proteome level analyses. Many software tools in this area combine efficient protein sequence filtering algorithms and spectral alignment algorithms to speed up database search. As a result, the performance of these tools heavily relies on the sensitivity and efficiency of their filtering algorithms. Here, we propose two efficient approximate spectrum‐based filtering algorithms for proteoform identification. We evaluated the performances of the proposed algorithms and four existing ones on simulated and real top‐down mass spectrometry data sets. Experiments showed that the proposed algorithms outperformed the existing ones for complex proteoform identification. In addition, combining the proposed filtering algorithms and mass graph alignment algorithms identified many proteoforms missed by ProSightPC in proteome‐level proteoform analyses. 相似文献